Alkaline proteases produced by Bacillus licheniformis RP1 grown on shrimp wastes: Application in chitin extraction, chicken feather-degradation and as a dehairing agent

The aim of this work was to study some biochemical characteristics of crude alkaline protease extracts from the viscera of goby (Zosterisessor ophiocephalus), thornback ray (Raja clavata), and scorpionfish (Scorpaena scrofa), and to investigate their applications in the deproteinization of shrimp wastes. At least four caseinolytic proteases bands were observed in zymogram of each enzyme preparation. The optimum pH for enzymatic extracts activities of Z. ophiocephalus, R. clavata, and S. scrofa were 8.0-9.0, 8.0, and 10.0, respectively. Interestingly, all the enzyme preparations were highly stable over a wide range of pH from 6.0 to 11.0. The optimum temperatures for enzyme activity were 50∘C for Z. ophiocephalus and R. clavata and 55∘C for S. scrofa crude alkaline proteases. Proteolytic enzymes showed high stability towards non-ionic surfactants (5% Tween 20, Tween 80, and Triton X-100). In addition, crude proteases of S. scrofa, R. clavata, and Z. ophiocephalus were found to be highly stable towards oxidizing agents, retaining 100%, 70%, and 66%, respectively, of their initial activity after incubation for 1 h in the presence of 1% sodium perborate. They were, however, highly affected by the anionic surfactant SDS. The crude alkaline proteases were tested for the deproteinization of shrimp waste in the preparation of chitin. All proteases were found to be effective in the deproteinization of shrimp waste. The protein removals after 3 h of hydrolysis at 45∘C with an enzyme/substrate ratio (E/S) of 10 were about 76%, 76%, and 80%, for Z. ophiocephalus, R. clavata, and S. scrofa crude proteases, respectively. These results suggest that enzymatic deproteinization of shrimp wastes by fish endogenous alkaline proteases could be applicable to the chitin production process.

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Study on the Fermentation Conditions and the Application in Feather Degradation of Keratinase Produced by Bacillus licheniformis

Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012) - Lecture Notes in Electrical Engineering ◽

10.1007/978-3-642-37916-1_10 ◽

2013 ◽

pp. 89-98

Author(s):

Yu Li ◽

Shuai Fan ◽

Sheng Chen ◽

Hao Er ◽

Jianjie Du ◽

...

Keyword(s):

Bacillus Licheniformis ◽

Feather Degradation ◽

Fermentation Conditions

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Rapid Conversion of Chicken Feather to Feather Meal Using Dimeric Keratinase from Bacillus licheniformis ER-15

Journal of Bioprocessing & Biotechniques ◽

10.4172/2155-9821.1000123 ◽

2012 ◽

Vol 02 (04) ◽

Cited By ~ 32

Author(s):

Ekta Tiwary

Keyword(s):

Bacillus Licheniformis ◽

Chicken Feather ◽

Feather Meal ◽

Rapid Conversion

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Screening and Characterisation of Keratin-Degrading Bacillus sp. from Feather Waste

Biotechnology Journal International ◽

10.9734/bji/2019/v23i130067 ◽

2019 ◽

pp. 1-12

Author(s):

M. T. Dada ◽

S. M. Wakil

Keyword(s):

Bacillus Subtilis ◽

Proteolytic Activity ◽

Bacillus Licheniformis ◽

Basal Medium ◽

Bacillus Species ◽

Bacillus Sp ◽

Feather Degradation ◽

Feather Waste ◽

Degrading Bacteria ◽

Significant Difference

Aim: This study focuses on the screening and characterisation of keratin-degrading Bacillus species from feather waste. Methods: Nine bacteria were isolated from feather waste obtained from a poultry layout at Egbeda local government secretariat, Ibadan, Nigeria. These bacteria were grown in basal medium with feather as primary source of carbon, nitrogen, sulfur and energy. Feather degrading bacteria were screened for both proteolytic activity and keratin degradation on skimmed milk agar and keratin azure medium respectively. They were also screened for their ability to degrade other keratin substrates such as hair and nail. Results: Three of the isolates with higher feather degradation levels also showed high proteolytic activity and release of azure dye. They were selected and identified phenotypically and genotypically using 16S rRNA sequencing as Bacillus licheniformis-K51, Bacillus subtilis-K50 and Bacillus sp.-K53. The bacteria were capable of degrading other keratin-containing substrates such as nail and hair. Bacillus subtilis-K50 and Bacillus licheniformis-K51 showed significant difference (P) in degradation among the three different keratin sources used yielding higher degradation with feather as keratin source with respective optical densities of 0.07 and 0.11 followed by hair and least in nails with optical densities of 0.05 and 0.07 respectively. Highest degradation of all the three keratin substrates was observed in Bacillus licheniformis-K51. Conclusion: The three isolated bacteria possess the ability to degrade keratin and utilize feather as keratin substrate. As a result, these can be considered as potential candidates for degradation and utilization of feather keratin.

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DEMINERALISASI DAN DEPROTEINASI KULIT UDANG SECARA KONTINYU PADA TAHAPAN EKSTRAKSI KITIN SECARA BIOLOGIS

Jurnal Sains dan Teknologi Indonesia ◽

10.29122/jsti.v12i1.843 ◽

2012 ◽

Vol 12 (1) ◽

Author(s):

Deden Rosid Waltam ◽

Heri Hermansyah ◽

Siswa Setyahadi

Keyword(s):

Protein Content ◽

Optimal Condition ◽

Bacillus Licheniformis ◽

Retention Time ◽

Lactobacillus Acidophilus ◽

Chemical Process ◽

Batch Fermentation ◽

Fermentation Process ◽

Alternative Technology ◽

Chitin Extraction

Chitin extraction in industry has been conducted by chemical process. The process has been known as a harsh treatment that badly affected to chitin quality, equipment and the environment. Since the last decade biologically chitin extractionhas more attracted attention. The biologically chitin extraction was conducted by batch fermentation or subsequent-batch fermentation. Continous demineralization and deproteinization is a new inovation on biologically chitin production technology.This system promises as an alternative technology for overcoming problems of batch fermentation process and chemical process. The objectives of the experiment was to obtain the optimal condition for continous deminineralizationand deproteinization for producing chitin from Panaeus vannamei shrimp shells. Lactobacillus acidophilus FNCC 116 and Bacillus licheniformis F11.1 was used for demineralization and deproteination process respectively. The results showed that the best condition for continuous demineralization was 6,5% glucose feed, with 16 hours retention time. For continuous deproteinization, the best condition was with 12 hours retention time. The process could remove 92.95% ash and 91.40% protein. The chitin, ash, and protein content of chitin product was 96.69%, 1.44% and 1,76% respectively.

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Bacillus sp. FPF-1 Produced Keratinase with High Potential for Chicken Feather Degradation

Molecules ◽

10.3390/molecules25071505 ◽

2020 ◽

Vol 25 (7) ◽

pp. 1505 ◽

Cited By ~ 6

Author(s):

Nonso E. Nnolim ◽

Anthony I. Okoh ◽

Uchechukwu U. Nwodo

Keyword(s):

Time Course ◽

Process Condition ◽

Agitation Speed ◽

Bacillus Sp ◽

Chicken Feather ◽

Feather Degradation ◽

Electron Microscopic ◽

16S Rdna Sequence ◽

Microscopic Imaging ◽

Chicken Feathers

Chicken feathers are predominantly composed of keratin; hence, valorizing the wastes becomes an imperative. In view of this, we isolated keratinase-producing bacteria and identified them through the 16S rDNA sequence. The process condition for keratinase activity was optimized, and electron micrography of the degradation timelines was determined. Keratinolytic bacteria were isolated and identified as Bacillus sp. FPF-1, Chryseobacterium sp. FPF-8, Brevibacillus sp. Nnolim-K2, Brevibacillus sp. FPF-12 and Brevibacillus sp. FSS-1; and their respective nucleotide sequences were deposited in GenBank, with the accession numbers MG214993, MG214994, MG214995, MG214996 and MG214999. The degree of feather degradation and keratinase concentration among the isolates ranged from 62.5 ± 2.12 to 86.0 ± 1.41(%) and 214.55 ± 5.14 to 440.01 ± 20.57 (U/mL), respectively. In the same vein, 0.1% (w/v) xylose, 0.5% (w/v) chicken feather, an initial fermentation pH of 5.0, fermentation temperature of 25 °C and an agitation speed of 150 rpm, respectively, served as the optimal physicochemical conditions for keratinase activity by Bacillus sp. FPF-1. The time course showed that Bacillus sp. FPF-1 yielded a keratinase concentration of 1698.18 ± 53.99(U/mL) at 120 h. The electron microscopic imaging showed completely structural dismemberment of intact chicken feather. Bacillus sp. FPF-1 holds great potential in the valorization of recalcitrant keratinous biomass from the agro sector into useful products.

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